KR20230169073A - Clinical Dosing Schedule for Inositol Phosphate Oligo(ethylene Glycol) Compounds - Google Patents

Abstract

The present invention relates to a dosing schedule for an inositol polyphosphate oligo(ethylene glycol) compound, or a pharmaceutically acceptable salt thereof, for use in the treatment or prevention of diseases associated with the formation and/or tissue exposure of calcium salt crystals. The drug compound is administered at weekly intervals.

Description

Clinical Dosing Schedule for Inositol Phosphate Oligo(ethylene Glycol) Compounds

The present invention relates to compounds and compositions for use in the treatment of conditions associated with calcification of tissue, particularly renal tissue, caused by deposition or exposure to calcium phosphate (CaP) and other calcium deposits.

This application claims priority from European application EP21153605, filed January 26, 2021, the entire contents of which are incorporated herein by reference.

WO2013045107 (A1) discloses for the first time the concept of using inositol polyphosphate oligo(ethylene glyco) derivatives as pharmaceuticals. Although it was initially thought to be an agent with significant potential to neutralize C. difficile toxin in the colonic lumen, subsequent analysis showed that it was initially disclosed in WO2017098047 (A1), US10624909 (B2) and US20200247837 (A1). Likewise, this compound was found to be very effective in reducing calcification when applied systemically.

All patent documents mentioned herein are incorporated by reference in their entirety.

The present inventors have examined the pharmacological profile of inositol phosphate oligo(ethylene glycol) derivatives in several studies and found that the compounds do not accumulate after repeated daily administration over 7 days (Schantl et al., Nature Communications 11, 721 (2020); https://doi.org/10.1038/s41467-019-14091-4).

The object of the present invention is to provide an advantageous dosing schedule for inositol polyphosphate oligo(ethylene glycol) derivatives. This object is achieved by the subject matter of the independent claims of the present specification.

Summary of the Invention

The present invention relates to administration schedules of inositol polyphosphate oligo(ethylene glycol) compounds, or pharmaceutically acceptable salts thereof, for use in the treatment or prevention of diseases associated with the formation and/or tissue exposure of calcium salt crystals. According to the invention, the inositol polyphosphate oligo(ethylene glycol) compound is administered at weekly or longer intervals.

Terms and Definitions

For purposes of interpreting this specification, the following definitions apply and, where appropriate, terms used in the singular include the plural and vice versa. If the definitions set forth below conflict with the documents incorporated herein by reference, the set forth definitions shall control.

As used herein, “comprising,” “having,” “containing,” “including,” and other similar forms and corresponding grammatical forms have equivalent meanings. It is intended to be open-ended in the sense that the items following one of these words are not a complete list of items or limited to only the items listed. For example, an item that “comprises” components A, B, and C may consist of only components A, B, and C (i.e., contains only components), or it may consist of components A, B, and C as well as one or more of the components A, B, and C. It may also contain other components. Accordingly, “comprises” and similar forms and grammatical equivalents are intended to include disclosure of embodiments of “consisting essentially of” or “consisting of” and I understand.

When a range of values is given, unless the context clearly indicates otherwise, the upper and lower limits of that range and each intermediate value between the other stated or intermediate values of that range shall be equal to one unit of the lower limit, subject to any limits specifically excluded from the stated range. It is understood that up to 1/10th is included within the scope of disclosure. If the stated range includes one or both of the two limits, the range excluding either or both of the included limits is also included in the disclosure.

In this document, “about” a value or parameter includes (and describes) variations on that value or parameter itself. For example, a description referring to “about X” includes description of “X.”

As used herein, including the appended claims, the singular forms "a", "or" and "the" include plural references unless the context clearly dictates otherwise.

In the context of this specification, the term oligo( ethylene glycol ) refers to oligomers of close chemical relatives such as ethylene glycol and oligo-propylene glycol and oligo-glycerol. The term “oligo” indicates the presence of one or more, especially 2 to 20, more particularly 2 to 12 monomers (-CH ₂ -CH ₂ -O-).

In the context of this specification, the term calcification relates to the formation of calcium deposits in affected tissues, particularly renal tissue or the cardiovascular system.

The term “weekly interval”, when applied to the administration of a compound, means that the compound is administered to the patient on day 1 of the dosing schedule, then on day 8, but not on any intermediate days.

DETAILED DESCRIPTION OF THE INVENTION

A first aspect of the invention relates to inositol polyphosphate oligo(ethylene glycol) compounds or pharmaceutically acceptable salts thereof for use in the treatment or prevention of diseases associated with formation and/or tissue exposure of calcium salt crystals, comprising: inositol The polyphosphate oligo(ethylene glycol) drug compound is administered at weekly intervals or at longer intervals exceeding one week.

In the broadest sense, diseases for which a weekly dosing schedule is provided are heart or cardiovascular diseases associated with the formation of calcium salt deposits, especially deposits consisting of calcium phosphate and/or oxalate.

In certain embodiments, the inositol polyphosphate oligo(ethylene glycol) compound, or pharmaceutically acceptable salt thereof, for use in accordance with the invention by weekly administration is represented by general formula (I):

(I)

where one or two or three X 's are oligo(ethylene glycol) and the remaining X's are OPO ₃ ^2- .

In certain embodiments, the inositol polyphosphate oligo(ethylene glycol) compound, or pharmaceutically acceptable salt thereof, is depicted by Formula I, wherein two Xs are oligo(ethylene glycol) and the remaining four Xs are OPO ₃ ^2- . This includes the bis-oligoethylene myo-inositol tetrakisphosphate compound represented by formula (I a), which is the compound used in the examples.

In certain embodiments, an inositol polyphosphate oligo(ethylene glycol) compound, or a pharmaceutically acceptable salt thereof, for use in accordance with the invention by weekly administration is represented by Formula I, wherein the three The remaining three Xs are OPO ₃ ^2- . The present inventors have previously shown that tris-oligoethylene myo-inositol trisphosphate compounds have comparable calcium salt precipitation prevention efficacy to bis-OEG compounds.

In certain embodiments, an inositol polyphosphate oligo(ethylene glycol) compound, or a pharmaceutically acceptable salt thereof, for use in accordance with the invention by weekly administration is represented by Formula I, wherein one X is oligo(ethylene glycol) and the other 5 X is OPO ₃ ^2- .

In certain embodiments, an inositol polyphosphate oligo(ethylene glycol) compound, or a pharmaceutically acceptable salt thereof, for use in accordance with the invention by weekly administration is represented by Formula I, wherein oligo(ethylene glycol) has Formula O— (CH ₂ -CH ₂ -O) _n CH ₃ , where n is selected from an integer between 2 and 20 , and in particular n is 2 to 12.

In a more specific embodiment, the inositol polyphosphate oligo(ethylene glycol) compound, or pharmaceutically acceptable salt thereof, for use in accordance with the invention by weekly administration is represented by Formula I, wherein the oligo(ethylene glycol) is of Formula O -(CH ₂ -CH ₂ -O) _n CH ₃ , and n is 2 .

In a very specific embodiment, the inositol polyphosphate oligo(ethylene glycol) compound, or pharmaceutically acceptable salt thereof, is depicted by Formula I a:

(I a)

In another specific embodiment, the inositol polyphosphate oligo(ethylene glycol) compound, or pharmaceutically acceptable salt thereof, is depicted by Formula I b:

(I b)

In another specific embodiment, the inositol polyphosphate oligo(ethylene glycol) compound, or pharmaceutically acceptable salt thereof, is depicted by Formula I c:

(I c)

In another specific embodiment, the inositol polyphosphate oligo(ethylene glycol) compound or pharmaceutically acceptable salt thereof is depicted by Formula I d:

(Id)

In another specific embodiment, the inositol polyphosphate oligo(ethylene glycol) compound, or pharmaceutically acceptable salt thereof, is depicted by Formula I e:

(Ie)

In certain embodiments, the compound is administered to human patients at a dose of 60 nmol/kg/day to 1700 nmol/kg/day.

The inventors calculated the dose based on the observed profile of Compound I a (see Examples and Figures) to be effective in the mouse model in the range of about 0.5 mg/kg/day to 15 mg/kg/day. Using well-known conversion figures (see: Guidance for Industry; Estimating the Maximum Safe Starting Dose in Initial Clinical Trials for Therapeutics in Adult Healthy Volunteers; U.S. Department of Health and Human Services Food and Drug Administration, Center for Drug Evaluation and Research (CDER), July 2005), the conversion factor for conversion from mouse to human dosing schedule is 0.08 and ranges from 40 to 1200 μg/kg/day. Using a molecular weight of (I a) of 698 g/mol (not counting counterions by weight) gives the above range.

In certain embodiments, the inositol polyphosphate oligo(ethylene glycol) compound is administered at a dose of 65 μg/kg/day to 1100 μg/kg/day in human subjects or 0.8 mg/kg/day to 14 mg/kg/day in mouse models. Delivered weekly at a dose between 90 nmol/kg/day and 1600 nmol/kg/day, equivalent to

In another specific embodiment, the inositol polyphosphate oligo(ethylene glycol) compound is administered at a dose of 170 nmol/kg/day to 2500 nmol/kg/day, equivalent to a dose of 120 μg/kg/day to 1750 μg/kg/day in a human subject. It is delivered weekly in doses.

In a more specific example, the inositol polyphosphate oligo(ethylene glycol) compound is administered at 170 to 600 nmol/kg/day, which is equivalent to 120 to 400 μg/kg/day of Compound (I a) at 1.5 to 5 mg/kg/day in a mouse model. Delivered weekly in doses between nmol/kg/day.

In certain embodiments, the inositol polyphosphate oligo(ethylene glycol) compound, or a pharmaceutically acceptable salt thereof, is administered at a dose that maintains plasma levels greater than 10 nm/mL, particularly plasma levels between 10 ng/mL and 50 μg/mL. is administered for use according to the invention at a dose that maintains a plasma level of 100 ng/mL to 5 μg/mL.

In the examples, the inositol polyphosphate oligo(ethylene glycol) compound was administered for weekly administration as a solution at a concentration of approximately 0.1 mL/10 g body weight (10 ml/kg) according to standard practice. The solution is adjusted to a neutral or slightly acidic pH in the pH range of 6 to 7.5. Sodium salt was used in the examples presented herein.

calcium deposits

The chemical composition of calcium crystal deposits associated with kidney and cardiovascular disease is well studied (Elliot, J Urology 100 (1968), 687-693; Xie et al., Cryst Growth Des. 2015 Jan 7; 15(1): 204 -211). In principle, treatment according to the invention can prevent or ameliorate any condition in which the deposition of calcium phosphate, oxalate and mixed calcium phosphate-oxalate crystals plays a role.

Calcium phosphate is found in various crystal forms in pathologic deposits in the human body; These include hydroxyapatite (hydroxylapatite, HA, Ca ₁₀ (PO ₄ ) ₆ (OH) ₂ ), brushite (CaHPO ₄ ^* 2 H ₂ O) and monetite. ) and various amorphous calcium phosphate salts.

Calcium oxalate in its pure form is found as mono(whewellite), di(weddellite) and trihydrate, and is often associated with other precipitates, primarily phosphate salts.

The results presented in the examples were obtained with a murine model of pseudoxanthoma elasticum (PXE), a prototype of the hereditary ectopic calcification disorder.

PXE affects the skin, eyes, and cardiovascular system due to inactivating mutations in the ABCC6 gene. In the examples shown below, we demonstrate the efficacy of inositol polyphosphate oligo(ethylene glycol) compound (I a) in inhibiting ectopic calcification in the Abcc6-/- mouse model of PXE.

In a prevention study, Abcc6-/- mice received 0.16, 0.8, 4, 20, or 100 mg/kg/day administered by subcutaneous implantation of an osmotic pump, as well as by subcutaneous injection three times weekly, at 6 weeks of age, when ectopic calcification begins. Treatment was with the compound at 4 mg/kg/day administered or 14, 4 and 0.8 mg/kg/day by subcutaneous injection once weekly. Mice were necropsied at 12 weeks of age. Histological examination and quantitative calcium analysis showed that mice administered the drug continuously for 6 weeks via an osmotic pump showed a complete response at 4 mg/kg/day and muzzle skin calcification at an effective dose of 0.8 mg/kg/day. showed a dose-dependent inhibition of Plasma concentrations of drug compounds were dose dependent and generally consistent throughout the treatment period for each dose. Subcutaneous injections three times a week and once a week (I a) also prevented calcification. In a treatment study, 12-week-old Abcc6-/- mice with evident calcification were administered INS-3001 at 4 mg/kg/day continuously for 12 weeks. Treatment according to the invention has been found to stabilize existing calcifications that were in progress at the start of treatment. These results demonstrate that the compounds and schedules according to the invention provide a promising approach to treating any ectopic calcification disorder.

The results presented herein facilitate the use of weekly treatment schedules for several diseases.

Diseases associated with calcium deposits that benefit from treatment according to the invention

“Renal fibrosis” or “tubulointerstitial fibrosis”, which involves the formation of calcium salt crystals and/or tissue exposure to wound healing after chronic injury due to exposure to calcium phosphate, calcium oxalate, or mixed CaP/CaOx deposits. Failure to do so results in thickening of the kidney tissue and the formation of scars. During this process, fibrotic matrix deposition continues unchecked, causing glomerulosclerosis, tubular atrophy, and interstitial fibrosis. Patients with the disease may experience severe abdominal pain (with bleeding or bleeding), swelling and discoloration of one or both legs, and may ultimately progress to chronic kidney disease.

“Renal inflammation” or “nephritis” is characterized by the recruitment of circulating monocytes, lymphocytes and neutrophils, as well as renal parenchymal cells, macrophages and dendritic cells. It is defined as a complex network of interactions between resident immune cells such as macrophages and dendritic cells. When stimulated, these cells activate special structures such as Toll-like receptors and Nod-like receptors (NLR). These receptors sense danger-related molecules and trigger key innate immune pathways, such as nuclear factor κB (NF-κB) and the NLRP3 inflammasome, resulting in metabolic reprogramming and phenotypic changes in immune and parenchymal cells. It can trigger the release of several inflammatory mediators that can cause severe and irreversible tissue damage and loss of function. In CKD, chronic inflammation can progressively reduce glomerular filtration rate (GFR), ultimately leading to kidney failure (end stage renal disease, ESRD).

Where kidney inflammation/nephritis is associated or caused by the interaction of calcium deposits with kidney tissue, treatment according to the invention is expected to improve or prevent the condition. The examples disclosed herein demonstrate that calcium phosphate deposition/exposure leads to upregulation of markers of inflammation and fibrosis in the kidney, and therefore all diseases consisting of inflammation and fibrosis of the kidney should benefit from treatment that inhibits deposit formation. do. Because inflammation and fibrosis are often multifactorial processes, each of these inflammatory diseases may have different causes that simultaneously cause inflammation and fibrosis. However, eliminating one causative factor is expected to improve the overall clinical picture.

“Glomerulonephritis” refers to a group of diseases characterized by inflammatory changes in the glomerular capillaries. Patients suffering from this disease may be accompanied by proteinuria, kidney dysfunction, fluid retention, high blood pressure, and edema. Glomerulonephritis may occur primarily as a kidney disease, but may also represent a systemic disease process. Glomerulonephritis associated with the formation and/or tissue exposure of calcium salt crystals is expected to benefit from treatment with the compounds described herein.

“Interstitial nephritis” refers to inflammation of the kidney interstitium that can be caused by an imbalance in calcium levels. Symptoms include increased urine output, hematuria, mental status changes, and edema. If this condition is associated with the formation and/or tissue exposure of calcium salt crystals, it is expected to benefit from treatment with the compounds described herein.

“Phosphate-induced renal fibrosis” may be associated with varying calcium and phosphate concentrations that cause precipitation in the renal tubules. See data presented in Figure 2: calcium > 2 or 5 mmol/L and phosphate > 5 or 7 mmol/L). It is important to note that even when detectable plasma phosphate levels are normal (i.e., in the absence of hyperphosphatemia), phosphate may be increased in renal tubular fluid (leading to calcium deposition). Elevated plasma phosphate is a late-stage consequence of end-stage renal disease, which occurs when kidney function is below the 20% threshold.

“Phosphate-induced chronic kidney disease” Phosphate-induced chronic kidney disease (CKD) can occur even when plasma phosphate levels are normal (i.e., in patients with early-stage chronic kidney disease). The cutoff values are calcium > 2 or 5 mmol/L and phosphate > 5 or 7 mmol/L in the renal tubules.

“Chronic kidney disease associated with hyperphosphatemia” is hyperphosphatemia that occurs in progressive chronic kidney disease (CKD) and occurs due to increased loss of glomerular filtration rate (GFR). This blocks tubular phosphate reabsorption, increases phosphate retention, reduces phosphate clearance, and impairs phosphate homeostasis (Sharon M. Moe, Prim Care. 2008 June; 35(2): 215-vi.). A useful cut-off value for characterizing patients expected to benefit from treatment according to the invention may be a plasma phosphate level greater than 1.46 mmol/Ls.

“Progression of chronic kidney disease” is divided into five stages, from stage 1 (mild damage, glomerular filtration rate above 90) to stage 5 (complete renal failure, glomerular filtration rate below 15). Current guidelines define the critical glomerular filtration rate for chronic kidney disease as less than 60 mL/min per 1.73 m ² for 3 months.

“Phosphate toxicity” refers to a condition in which phosphate homeostasis is impaired due to impaired regulation of renal phosphate excretion and reabsorption, which can cause serious damage to renal tissue (Razzaque, Clin Sci (Lond). 2011 Feb; 120( 3): 91-97). To the extent that such conditions lead to formation of crystals and/or tissue damage associated with tissue exposure to and/or deposition of crystals, which can be avoided by treatment according to the invention, the treatment is suitable for patients suffering from that condition.

“Hyperphosphaturia” or “phosphaturia” refers to the condition of high phosphate levels in the urine. To the extent that this condition causes the formation of crystals and/or tissue damage associated with tissue exposure to and/or deposition of the crystals, which can be avoided by treatment according to the invention, the treatment is intended for use in patients suffering from that condition. It is instructed.

“Hyperphosphatemia” refers to a condition in which blood phosphate levels are elevated (> 4.5 mg/dL; > 1.46 mmol/L). To the extent that this condition causes the formation of crystals and/or tissue damage associated with tissue exposure to and/or deposition of the crystals, which can be avoided by treatment according to the invention, the treatment is intended for use in patients suffering from that condition. It is instructed.

“Hyper-FGF23-emia” refers to decreased serum phosphate levels and increased fractional phosphate excretion due to elevated fibroblast growth factor (FGF)-23 levels.

“Vascular calcification” refers to the pathological deposition of minerals in the vascular system, often observed in patients suffering from CDK or diabetes. Elevated calcium and/or phosphate levels may be the result of metabolic dysregulation due to diabetes, dyslipidemia, oxidative stress, uremia, and hyperphosphatemia, leading to the formation of osteoblast-like cells, Leads to the formation of calcific deposits and stiffness.

“Coronary artery disease” or “atherosclerotic heart disease” refers to the buildup of plaque in the damaged lining of the coronary arteries. Factors such as inflammatory cells, lipoproteins, and calcium adhere to the plaque, causing further narrowing. If this disease progresses, it can ultimately lead to myocardial infarction or stroke.

“Vascular stiffening” refers to the hardening of artery walls due to calcification. Vascular stiffening consists of a decrease in the elasticity of blood vessels and an increase in pulse wave pressure.

“Valve calcification,” especially aortic valve calcification, refers to the calcification of valves, especially the aortic and mitral valves, while simultaneously causing ECM decomposition, fibrosis, lipid accumulation, and neo-angiogenesis in valve tissue. It refers to a disturbance in the active regulation of normal homeostatic processes and hemodynamic changes.

“Nephrocalcinosis” refers to the deposition of calcium salts in the kidney parenchyma, especially in the medulla (medullary nephrocalcinosis) or cortex (cortical nephrocalcinosis) of the kidney. It says what happens.

“Calcinosis cutis” refers to the deposition of calcium phosphate deposits on the skin, especially in the extremities. When the melting point of calcium and phosphate is exceeded, precipitation of calcium salts and deposition as amorphous hydroxyapatite occurs.

“Kidney stones,” “renal calculi,” “nephrolithiasis,” and “urolithiasis” refer to supersaturation of urine (hypercalciuria) due to accumulation of minerals in kidney tissue. .

“Chondrocalcinosis” refers to the accumulation of calcium phosphate in the joints.

Based on the examples described herein, specific diseases for which therapeutic effects are expected to be achieved by administering the compounds described herein further include aortic valve stenosis, peripheral artery disease, and cerebral calcification.

In certain embodiments, the diseases associated with calcification that are addressed by the weekly dosing regimen according to the present invention include myocardial infarction, cardiovascular death, progression of chronic kidney disease, peripheral artery disease, critical limb ischemia, calcinosis, and systemic arterial calcification of infancy. , aortic stenosis, atherosclerosis, pseudoxanthoma elasticum, cardiac hypertrophy, heart failure, arrhythmias, aneurysms, valvular stenosis, aortic regurgitation, mitral stenosis, mitral regurgitation, stroke, and cognitive decline, each of which increases calcium in the affected tissues. It is related to salt deposition.

The data presented in Example 1 show that the use of idiopathic calcium nephrolithiasis/idiopathic calcium nephrolithiasis, especially calcium phosphate nephrolithiasis consisting primarily of calcium phosphate, calcium oxalate or mixtures thereof, may reduce calcium phosphate crystal formation or prevent the treatment of calcium nephrolithiasis. confirms the utility of the compounds disclosed herein to be effective in inhibiting

In certain embodiments, inositol polyphosphate oligo(ethylene glycol) compounds, or pharmaceutically acceptable salts thereof, for use in the treatment or prevention of the diseases set forth above are described by general formula (I), wherein one or two or three It is oligo(ethylene glycol) and the remaining X is OPO ₃ ^2- .

(I)

It is clear to those skilled in the art that when dissolved in aqueous media at physiological pH, the compounds mentioned herein are anionic and will carry cations.

In certain embodiments, two ^of the _inositol substituents

Examples of compounds that may advantageously be used in the treatment according to the invention include the general formulas I-1, I-2, I-3 and I-4, in each case ^as specified herein wherein is oligo(ethylene glycol):

(I-1) (I-2)

(I-3) (I-4)

In one or more specific embodiments thereof, the scaffold has the myo-inositol and oligo(ethylene glycol) substituents at positions 4 and 6, and the remaining substituents are phosphate. In an even more specific embodiment, the oligo(ethylene glycol) substituent is O-(CH ₂ -CH ₂ O) ₂ -CH ₃ .

In another specific embodiment, one ^of the _inositol substituents In one or more specific embodiments thereof, the scaffold is wherein the myo-inositol and oligo(ethylene glycol) substituents are at positions 4 or 6, especially at position 6, and the remaining substituents are phosphate. In an even more specific example, the oligo(ethylene glycol) substituent is O-(CH ₂ -CH ₂ O) ₂ -CH ₃ .

In another specific example, three of _the ^inositol substituents Examples thereof include general formulas I-5 and I-6 where R ¹ is oligo(ethylene glycol) as specified herein:

(I-5) (I-6),

In certain embodiments, the oligo(ethylene glycol) substituent of the inositol polyphosphate oligo(ethylene glycol) compound provided for use in accordance with the invention herein, or a pharmaceutically acceptable salt thereof, has the formula O-(CH2-CH2-O)nCH ₃ where n is selected from an integer between 2 and 20, and in particular n is 2 to 12. Various parameters of the compound's physiological activity, pharmacological parameters and manufacturing aspects influence the optimal n value.

In certain embodiments, the oligo(ethylene glycol) substituent of the inositol polyphosphate oligo(ethylene glycol) compound provided for use in accordance with the invention herein, or a pharmaceutically acceptable salt thereof, has the formula O—(CH ₂ -CH ₂ -O ) _n CH ₃ , where n is 2. The examples demonstrate certain advantages of (OEG ₂ ) ₂ -IP4(Ia), where n is 2.

Certain examples of compounds for use as specified herein are illustrated by any of the following formulas:

(I a)

(I b)

(I c)

Certain examples of compounds for use as specified herein are illustrated by any of the following formulas:

(I d)

(I e)

Other examples of compounds for use in accordance with aspects of the invention include:

(I f)

(I g)

Any of the inositol polyphosphate oligo-alkyl ether compounds or pharmaceutically acceptable salts thereof are provided for use in the treatment or prevention of diseases associated with the formation of calcium phosphate salts or other solid deposits in the body, particularly in kidney tissue.

Administration of a compound with two oligo(ethylene glycol) moieties attached to a myo-inositol tetrakisphosphate scaffold (I a) is demonstrated by the data provided in Example 1 and is effective in reducing calcium phosphate-based Randall's plaques. Treatment or prevention of diseases as described herein associated with precipitation of calcium phosphate solids, as exemplified by the growth of calcium oxalate kidney stones (based on Randall's plaque) and mixed phosphate-oxalate calcium phosphate deposits or predominantly oxalate. However, it is particularly advantageous for preventing or treating deposits originating from calcium phosphate nuclei.

Bipegylated compounds such as (OEG2) ₂ -IP4 also have a protective effect with respect to mixed precipitates (CaP+CaOx).

Those skilled in the art recognize that any of the specifically mentioned drug compounds mentioned herein may exist as pharmaceutically acceptable salts of said drugs. Pharmaceutically acceptable salts include ionized drugs and oppositely charged counterions. Non-limiting examples of pharmaceutically acceptable cationic salt forms include aluminum, benzathine, calcium, ethylene diamine, lysine, magnesium, meglumine, potassium, procaine, sodium, tromethamine, and zinc.

Manufacturing method and treatment method according to the present invention

In a further embodiment the invention provides for weekly administration, diseases associated with the formation of calcium salt deposits or calcium salt crystals, in particular calcification of renal tissue, nephritis, in particular interstitial nephritis, glomerulonephritis, phosphate-induced chronic kidney disease, hyperphosphatemia. Of medicaments for the treatment or prevention of diseases selected from chronic kidney disease, progression of chronic kidney disease, phosphate toxicity, hyperphosphatemia, hyperphosphatemia and/or hyper-FGF23-emia, specifically selected from calcification of renal fibrosis. It further includes the use of the inositol polyphosphate oligo(ethylene glycol) compound, or a pharmaceutically acceptable salt thereof, as specified above for use in a manufacturing process.

The compounds of the present invention are useful in treating diseases selected from the group consisting of diseases associated with the formation of calcium salt deposits or calcium salt crystals, vascular calcification, coronary artery disease, vascular sclerosis, valvular calcification, renal calciuria, dermatometriosis, nephrolithiasis and chondrocalcinosis. Similarly provided is for use in a method of preparing a medicament for treatment or prophylaxis.

Similarly, the present invention covers diseases associated with the formation of calcium salt deposits or calcium salt crystals, in particular calcification of renal tissue, cases associated with renal inflammation, in particular calcification of renal tissue, nephritis, in particular interstitial nephritis, glomerulonephritis, Phosphate-induced renal fibrosis, phosphate-induced chronic kidney disease, chronic kidney disease associated with hyperphosphatemia, progression of chronic kidney disease, phosphate toxicity, hyperphosphatemia, hyperphosphatemia and/or hyper-FGF23-emia, specifically renal fibrosis It includes a method of treating a patient diagnosed with a disease selected from. The method involves administering to a patient an effective amount of an inositol polyphosphate oligo(ethylene glycol) compound, or a pharmaceutically acceptable salt thereof, as detailed herein.

Pharmaceutical Compositions and Administration

Another aspect of the present invention relates to a pharmaceutical composition comprising a compound of the present invention or a pharmaceutically acceptable salt thereof. In another embodiment, the composition includes at least two pharmaceutically acceptable carriers as described herein.

In certain embodiments of the invention, the compounds of the invention are generally formulated in pharmaceutical dosage forms to provide easily controllable doses of the drug and to provide patients with an elegant and easily handled product.

In certain embodiments, pharmaceutical compositions for use in accordance with the invention are formulated for administration by intradermal or subcutaneous injection.

The dosing regimen for the compounds of the present invention depends on the pharmacodynamic properties of the specific agent and its mode and route of administration; Recipient's species, age, gender, health, medical condition and weight; nature and severity of symptoms; type of combination treatment; frequency of treatment; It will depend on known factors such as the route of administration, the patient's renal and hepatic function, and the desired effect. In certain embodiments, the compounds of the invention may be administered in a single daily dose or the total daily dose may be administered in two, three, or four divided doses per day.

Pharmaceutical compositions for use according to the invention may undergo customary pharmaceutical manipulations such as sterilization and/or may contain customary inert diluents or buffers as well as auxiliaries such as preservatives, stabilizers, surfactants and buffers, etc. . They can be produced by standard processes, for example conventional mixing, dissolving or freeze-drying processes. Many of these procedures and methods for preparing pharmaceutical compositions are known in the art, see, for example, L. Lachman et al. See The Theory and Practice of Industrial Pharmacy, 4th Ed, 2013 (ISBN 8123922892).

The present invention further includes the following items:

Item 1: An inositol polyphosphate oligo(ethylene glycol) compound or a pharmaceutically acceptable salt thereof for use in the treatment or prevention of diseases associated with the formation and/or tissue exposure of calcium salt crystals, comprising:

Inositol polyphosphate oligo(ethylene glycol) compound is administered at weekly intervals .

Item 2: Inositol polyphosphate oligo(ethylene glycol) compound or a pharmaceutically acceptable salt thereof for use according to item 1, wherein the inositol polyphosphate oligo(ethylene glycol) compound is represented by the general formula (I):

(I), where one or two or three Xs are oligo(ethylene glycol) and the remaining Xs are OPO ₃ ^2- .

Item 3: An inositol polyphosphate oligo(ethylene glycol) compound or a pharmaceutically acceptable salt thereof for use according to item 2, wherein two Xs are oligo(ethylene glycol) and the remaining four Xs are OPO ₃ ^2- .

Item 4: An inositol polyphosphate oligo ₍ ethylene glycol) compound or ^a pharmaceutically acceptable salt thereof for use according to item 2, wherein three

Item 5: An inositol polyphosphate oligo ₍ ethylene glycol) compound ^or a pharmaceutically acceptable salt thereof for use according to item 2, wherein one

Item 6: Item 2 to Fehler! Inositol polyphosphate oligo(ethylene glycol) compound or a pharmaceutically acceptable salt thereof for use according to Verweisquelle konnte nicht gefunden werden , wherein oligo(ethylene glycol) has the formula O-(CH ₂ -CH ₂ -O) _n CH ₃ where n is selected from an integer of 2 to 20 , and in particular n is 2 to 12.

Item 7: Item Fehler! An inositol polyphosphate oligo(ethylene glycol) compound or a pharmaceutically acceptable salt thereof for use according to Verweisquelle konnte nicht gefunden werden , where n is 2 .

Item 8: An inositol polyphosphate oligo(ethylene glycol) compound or a pharmaceutically acceptable salt thereof for use according to item 3, wherein the compound is represented by one of the following formulas:

(I a)

(I b)

(I c)

In particular, the compound herein is described by formula Ia.

Item 9: An inositol polyphosphate oligo(ethylene glycol) compound or a pharmaceutically acceptable salt thereof for use according to item 5, wherein the compound is represented by one of the following formulas:

(Id)

(Ie)

Item 10: An inositol polyphosphate oligo(ethylene glycol) compound or a pharmaceutically acceptable salt thereof for use according to any of the preceding items, wherein the compound has a dose of 60 nmol/kg/day to 2500 nmol/kg/day in human patients. With a capacity between

In particular at doses between 90 nmol/kg/day and 1600 nmol/kg/day,

More particularly administered at a dose between 170 nmol/kg/day and 600 nmol/kg/day,

This is equivalent to a dose of 40 to 1200 μg/kg/day, especially a dose of 65 μg/kg/day to 1100, more particularly a dose of 120 to 400 μg/kg/day (using compound (I a) and the counter ion weight not calculated, MW 698 gmol ^-1 ).

Item 11: An inositol polyphosphate oligo(ethylene glycol) compound or a pharmaceutically acceptable salt thereof for use according to any of the preceding items, wherein the compound is administered at a dose that maintains plasma levels exceeding 10 ng/mL in a human patient. , especially at a dose that maintains a plasma level between 10 ng/mL and 50 μg/mL, and more particularly at a dose that maintains a plasma level between 100 ng/mL and 5 μg/mL.

Item 12: An inositol polyphosphate oligo(ethylene glycol) compound or a pharmaceutically acceptable salt thereof for use according to any of the preceding items, wherein the compound or salt thereof is provided for the treatment or prevention of a disease selected from :

- Renal fibrosis, especially when associated with calcification of renal tissue,

- Kidney inflammation, especially when associated with calcification of kidney tissue,

- nephritis,

- Interstitial nephritis,

- Glomerulonephritis,

- Renal fibrosis due to phosphate,

- Phosphate-induced chronic kidney disease,

- Chronic kidney disease associated with hyperphosphatemia,

- progression of chronic kidney disease,

- Phosphate toxicity,

- Hyperphosphatemia

- vascular calcification,

- hyperphosphatemia, and/or

- Hyper-FGF23-emia.

Item 13: An inositol polyphosphate oligo(ethylene glycol) compound or a pharmaceutically acceptable salt thereof for use according to any of the preceding items, wherein the disease is associated with the formation of calcium phosphate salts or precipitates.

Surrender 14: An inositol polyphosphate oligo(ethylene glycol) compound or a pharmaceutically acceptable salt thereof for use according to any of the preceding clauses, wherein the disease is caused by the formation of calcium oxalate salts or precipitates and/or mixed calcium phosphate oxalate It is a disease associated with the formation of late deposits.

The invention is further illustrated by the following examples and drawings, from which further embodiments and advantages may be derived. These examples are intended to illustrate the invention but are not intended to limit its scope.

Figure 1 shows histopathological examination of vibrissae calcifications in the snout skin of different groups of Abcc6 ^-/- mice. (A), Prevention study. All mice were analyzed at 12 weeks of age. von Kossa staining revealed strong calcification in the dermal sheath of the whiskers in untreated Abcc6 ^−/− mice. Abcc6 ^-/- mice receiving (I a) via osmotic pump at 4 mg/kg/day had little calcification in the whiskers.
Abcc6 ^−/− mice receiving (I a) via osmotic pump at 0.16 mg/kg/day had no effect on calcification in the whiskers. Osmotic pump delivery of (I a) at 0.8 mg/kg/day resulted in an obvious but significant reduction in calcification. Subcutaneous injections of (I a) at different doses and injection frequencies all resulted in reduced calcification. (B), Reversal study. All mice were analyzed at 24 weeks of age. von Kossa staining revealed extensive whisker calcification in untreated Abcc6 ^−/− mice (I a) and treatment prevented further calcification, which stabilized upon treatment. Arrows indicate ectopic calcifications. Scale bar, 0.4 mm.
Figure 2 shows chemical analysis of calcium in snout skin biopsies and (I a) plasma concentrations of INS-3001 in mice receiving administration. (A), Calcium content of muzzle skin including whiskers. Plasma concentrations from 6–12 mice per group (B), (I a). 5 mice per group. For mice implanted with osmotic pumps, blood was collected before sacrifice or 30 min after subcutaneous injection. Overall, plasma concentrations were relatively well adjusted depending on the administered dose despite variability between different groups. The variability may be due to the fact that a single sampling time point was chosen around the expected T _max of 30 minutes, resulting in greater variability than if a complete pharmacokinetic profile had been obtained. * P < 0.05, ** P < 0.01 compared with untreated 12-week-old Abcc6 ^−/− mice; +P < 0.01 compared with untreated WT mice. Data were expressed as mean ± SD. nd, not decided.
Figure 3 shows plasma concentrations of (I a) in mice receiving continuous (I a) administration via osmotic pump. Blood was collected 2 days, 3 weeks, and 6 weeks after subcutaneous implantation of an osmotic pump filled with (I a) solution to deliver (I a) at 0.16, 0.8, 4, 20, and 100 mg/kg/day. 5 mice per group. * P < 0.05, ** P < 0.01 compared with untreated Abcc6 ^−/− mice. Data were expressed as mean ± SD.

Example

Materials and Methods

Abcc6 knockout mice (Abcc6 ^−/− ) on a C57BL/6J background were generated as previously described (JF Klement et al., Mol Cell Biol 2005, 25, 8299). C57BL/6J mice from The Jackson Laboratory (Bar Harbor, ME) were used as wild-type controls (WT). In a prevention study, 6-week-old Abcc6 ^−/− mice were administered (I a) by periodic subcutaneous injection or continuous administration via a subcutaneous ALZET osmotic pump model 2006 (DURECT Corp, Cupertino, CA). In a reversal study, 12-week-old Abcc6 ^−/− mice were administered (I a) at 4 mg/kg/day via osmotic pump for 12 weeks. All mice were maintained on a standard diet and sacrificed at 12 weeks of age 6 weeks after the start of treatment (prevention study) or at 24 weeks of age 12 weeks after the start of treatment (reversal study). All protocols were approved by the Institutional Animal Care and Use Committee at Thomas Jefferson University.

Snout skin biopsies (left) from euthanized mice were collected, fixed in 10% phosphate-buffered formalin, and embedded in paraffin. Paraffin sections (6 μm) were stained with Hematoxylin and Eosin or von Kossa using standard procedures. To quantify mineral deposition, rostral skin biopsies (right) were harvested and decalcified with 1.0 M HCl for 48 h at room temperature. Solubilized calcium was then measured by colorimetric analysis (J. Huang et al., J Invest Dermatol 2019, 139, 1254; Q. Li, J. Huang et al., J Invest Dermatol 2019, 139, 360). Values were normalized to tissue weight. Whole blood was collected by retro-orbital bleeding or cardiac puncture. For mice injected subcutaneously, whole blood was collected 30 min after injection (I a). Whole blood was collected from mice receiving (I a) 2 days, 3 weeks, and 6 weeks after initiation of treatment. The concentration of (I a) in heparin plasma was analyzed by ultra-performance liquid chromatography as previously described (A. E. Schantl et al., Nat Commun 2020, 11, 721). Data were analyzed using the Kruskal-Wallis nonparametric test. Statistical significance was reached at P < 0.05.

result

Example 1: Prevention Study: Young Abcc6 ^-/- Continuous delivery of (I a) to mice

Abcc6 ^−/− mice were administered (I a) by subcutaneous implantation of an osmotic pump at doses of 0.16, 0.8, 4, 20, and 100 mg/kg/day. Untreated wild-type and Abcc6 ^−/− mice served as negative and positive controls for ectopic calcification. Treatment was started at 6 weeks of age, the earliest stage of ectopic calcification in Abcc6 ^−/− mice. The extent of ectopic calcification after 6 weeks of treatment was analyzed in two independent assays measuring calcification in the calcification sheath of the whiskers of the rostral skin, which is an early and reliable biomarker in this PXE mouse model. A piece of snout skin was processed for semiquantitative histological evaluation. von Kossa staining, a calcium-specific stain, revealed strong calcification in the coronal sheath of the whiskers in untreated Abcc6 ^−/− mice at 12 weeks of age. Abcc6 ^-/- mice receiving (I a) at concentrations above 4 mg/kg/day showed little calcification in the snout skin. There is no dose correlation with (I a) in the range of 4-100 mg/kg/day, which means that the lowest dose of 4 mg/kg/day completely prevents ectopic calcification to the same extent as the highest dose of 100 mg/kg/day. This is because (almost 100% suppression). Significantly reduced calcification was observed at 0.8 mg/kg/day, in contrast 0.16 mg/kg/day had no effect on prevention of rostral skin calcification (Figure 1A). The therapeutic effect of (I a) was demonstrated by the calcium content in different rostral skin biopsies measured using quantitative chemical analysis (Figure 2a). The results showed that the calcium levels of Abcc6 ^−/− mice treated with (I a) above 4 mg/kg/day were similar to those of WT mice negative for ectopic calcification ( Fig. 2A ). Partial inhibition of ectopic calcification at 0.8 mg/kg/day was confirmed by calcium analysis corresponding to 59% inhibition (Figure 2A), thus confirming 0.8 mg/kg/day as the minimum active dose.

(I a) was not detected in the plasma of untreated Abcc6 ^−/− mice. Administration of (I a) at doses ranging from 0.16 to 100 mg/kg/day resulted in dose-dependent plasma concentrations (Figure 2b). Additionally, the level of (I a) remained almost constant at 2 days, 3 weeks, and 6 weeks after implantation of the osmotic pump (Figure 3), which was attributed to the nature of consistent and sustained release of (I a) through the osmotic pump. do.

Example 2: Prevention Study: Young Abcc6 ^-/- Intermittent subcutaneous injection of (I a) in mice.

When delivered continuously at 4 mg/kg/day by osmotic pump, (I a) completely prevented ectopic calcification in the snout skin of Abcc6 ^−/− mice. We next investigated whether periodic bolus subcutaneous injections of (I a) at the same dose of 4 mg/kg/day would have similar therapeutic benefits. Six-week-old Abcc6 ^−/− mice were administered (I a) by subcutaneous injection three times per week, equivalent to 4 mg/kg/day. These mice showed significantly reduced calcification in the snout skin 6 weeks after starting treatment, but residual calcification was still evident (Figure 1A), corresponding to 63% inhibition (Figure 2A). Therefore, subcutaneous injection of (I a) is less effective than continuous delivery at the same dose of 4 mg/kg/day for inhibition of ectopic calcification.

To evaluate the inhibitory effect of (I a), three additional dose levels of (I a) were tested in weekly subcutaneous injections over a period of 6 weeks (Figure 1a). The efficacy of weekly injections at 14 mg/kg/day was similar to 4 mg/kg/day administered three times weekly, suggesting that transient increases in plasma levels of (I a) may be beneficial. Abcc6 ^-/- mice receiving (I a) injections at a frequency of 4 mg/kg/day per week had significantly reduced calcification, similar to the effect when the same dose was injected at a frequency of three times per week. Weekly injections of (I a) at a low dose of 0.8 mg/kg/day also significantly reduced ectopic calcification, suggesting that (I a) is a potent inhibitor of ectopic calcification with pharmacological activity maintained over several days. (I a) Semiquantitative histological evaluation of the treatment was consistent with the calcium content of the snout skin measured by chemical analysis (Figure 2a).

It has been reported that the T _max of (I a) in plasma after subcutaneous administration to Sprague Dawley rats is about 30 minutes after injection. Therefore, the present inventors measured the concentration of (I a) in plasma 30 minutes after injection in Abcc6 ^-/- mice. did. At 30 minutes after injection, (I a) levels in plasma were significantly higher than those in the pump-administered group, as expected for bolus administration (Figure 2B). Prevention of (I a)-mediated ectopic calcification was more effective with continuous delivery reaching steady-state plasma concentrations, whereas bolus injections resulted in a transient increase in (I a) concentrations but were still significantly potent when administered periodically. .

Example 3: Treatment Study: Aged Abcc6 ^-/- Serial delivery of (I a) in mice

In the treatment study, Abcc6 ^−/− mice were aged up to 12 weeks, and ectopic calcification was extensive in the snout skin, including the whiskers ( Fig. 1B ). (I a) was administered at 4 mg/kg/day by osmotic pump, a dose and route of delivery that resulted in a complete response in young mice (Figure 1A). The degree of ectopic calcification was assessed after 12 weeks. (I a) Histopathology and calcium analysis of the snout skin of treated mice showed calcification similar to 12-week-old Abcc6 ^−/− control mice (Figs. 1, 2a). These results demonstrated that (I a) stabilized existing calcifications and stopped further growth of calcifications for a total of 12 weeks.

conclusion

Our results show that treatment with (I a) prevents rostral skin calcification in Abcc6 ^−/− mice with a concomitant increase in plasma levels, and that drug therapy with (I a) prevents PXE, a disease for which we currently lack prevention or treatment options. and that it may provide a promising treatment strategy for spontaneous calcification in GACI type 2. The mechanism of action of (I a) as a new calcification inhibitor is not through chelation of calcium, but rather through coating of calcification, preventing further mineralization. This mode of action is similar to that of the natural compound IP6, which was recently reported to halt, but not regress, the progression of cardiovascular calcification in dialysis patients after one year of treatment. (I a) arrested further mineral deposition at the point where treatment was initiated, suggesting that if (I a) is used to treat patients with PXE, it should be initiated as soon as the diagnosis is made. Unlike chaperone-based allele-specific therapy, (I a) has the potential to treat PXE regardless of the mutation type of ABCC6. Compared with pyrophosphate and bisphosphonate, (I a) has high subcutaneous bioavailability, good local tolerance, high efficacy and no anti-osteoclast activity.

Claims (6)

Inositol polyphosphate oligo(ethylene glycol) compounds described by compounds selected from any of formulas Ia, Ib, Ic for use in the treatment or prevention of diseases associated with the formation and/or tissue exposure of calcium salt crystals:
(I a)
(I b)
(I c)
or as a pharmaceutically acceptable salt of the compound,
The inositol polyphosphate oligo(ethylene glycol) compound is administered at one-week intervals,
Inositol polyphosphate oligo(ethylene glycol) compound or a pharmaceutically acceptable salt thereof. According to paragraph 1,
The compound is described by formula Ia,
Inositol polyphosphate oligo(ethylene glycol) compound or a pharmaceutically acceptable salt thereof. According to claim 1 or 2,
The disease associated with the formation and/or tissue exposure of calcium salt crystals is vascular calcification,
Inositol polyphosphate oligo(ethylene glycol) compound or a pharmaceutically acceptable salt thereof. According to claim 1 or 2,
The disease associated with the formation and/or tissue exposure of calcium salt crystals is a disease selected from the group consisting of an inositol polyphosphate oligo(ethylene glycol) compound or a pharmaceutically acceptable salt thereof:
- Renal fibrosis, especially when associated with calcification of kidney tissue,
- renal inflammation, especially when associated with calcification of kidney tissue,
- nephritis,
- interstitial nephritis,
- glomerulonephritis,
- Phosphate-induced renal fibrosis,
- Phosphate-induced chronic kidney disease,
- Chronic kidney disease associated with hyperphosphatemia,
- progression of chronic kidney disease,
- phosphate toxicity,
- hyperphosphaturia,
- hyperphosphatemia, and/or
- Hyper-FGF23-emia. According to any one of claims 1 to 4,
The disease is a disease associated with the formation of calcium phosphate salts or deposits,
Inositol polyphosphate oligo(ethylene glycol) compound or a pharmaceutically acceptable salt thereof. According to any one of claims 1 to 5,
Said related disease is a disease associated with the formation of calcium oxalate salts or precipitates and/or the formation of mixed calcium phosphate oxalate precipitates.
Inositol polyphosphate oligo(ethylene glycol) compound or a pharmaceutically acceptable salt thereof.